Franco Alhaique

Hyaluronic acid and dextran-based semi-IPN hydrogels as biomaterials for bioprinting.

Bioprinting is a recent technology in tissue engineering used for the design of porous constructs through layer-by-layer deposition of cell-laden material. This technology would benefit from new biomaterials that can fulfill specific requirements for the fabrication of well-defined 3D constructs, such as the preservation of cell viability and adequate mechanical properties. We evaluated the suitability of a novel semi-interpenetrating network (semi-IPN), based on hyaluronic acid and hydroxyethyl-methacrylate-derivatized dextran (dex-HEMA), to form 3D hydrogel bioprinted constructs. The rheological properties of the solutions allowed proper handling during bioprinting, whereas photopolymerization led to stable constructs of which their mechanical properties matched the wide range of mechanical strengths of natural tissues. Importantly, excellent viability was observed for encapsulated chondrocytes. The results demonstrate the suitability of hyaluronic acid/dex-HEMA semi-IPNs to manufacture bioprinted constructs for tissue engineering.

Modulation of caffeine skin delivery by carrier design: liposomes versus permeation enhancers

Abstract Delivery systems for caffeine have been designed for two different purposes: (1) enhancing drug permeation through the skin for systemic delivery and (2) accumulating drug reservoir in the skin for local delivery. Caffeine exhibited concentration-dependent growth inhibition of normal and psoriatic human fibroblasts, as well as 3T3 mouse fibroblasts. High flux of caffeine through the skin was obtained from an aqueous solution containing an enhancing mixture of 20% Transcutol and 10% oleic acid. The presence of the enhancers resulted in caffeine flux 40 times greater than in their absence. The high flux of caffeine through the skin in vitro which was obtained using the enhancing composition was shown to be parallel to increased serum concentrations of drug in rats in vivo. Application of caffeine in aqueous solution containing enhancing mixture resulted in high serum concentrations of 50–60 μ ml after 1 h, which remained high for at least 12 h following. The greatest caffeine accumulation in the skin was measured from small liposomal vesicles, 2260 solμg cm 2 , this being 3 times greater than from aqueous solution containing enhancers, the system which exhibited the second largest accumulation of drug in the skin. Using quantitative skin autoradiography, it was found that after 24 h, the greatest concentration of caffeine (280 μg g tissue) was localized in the epidermis and the lowest amount (50 μg g tissue) in the dermis. In addition, a relatively high concentration of caffeine was found in the appendages.

Preparation and properties of new unilamellar non-ionic/ionic surfactant vesicles

Abstract Non-ionic surfactant vesicles (NSVs) were prepared from polysorbate 20 and cholesterol by means of two different methods: by direct sonication of an aqueous dispersion of the various components (bulk) or by solubilization of the components, evaporation of the organic solvent to form a film inside the vessel used for the preparation and then by sonication (film). The influence of the preparation technique on the properties of the obtained structures was studied. Vesicles with bigger dimensions and higher entrapment efficiency were obtained when sonication was carried out after the film formation. Vesicle formation in the presence of ionic surfactants was investigated in order to evaluate the effect of charged components on vesicle dimensions, entrapment efficiency and stability. Dimethyldioctadecylammonium bromide (DDOA) and cetylpyridinium chloride (CPy) were used to introduce a positive charge in the vesicle structure, while dicetylphosphate (DCP) was used for a negative charge. Better resistance to osmotic stress and higher entrapment efficiency values were obtained with vesicles containing DCP and CPy.

Non-ionic surfactant vesicles in pulmonary glucocorticoid delivery: Characterization and interaction with human lung fibroblasts

Non-ionic surfactant vesicles (NSVs) were proposed for the pulmonary delivery of glucocorticoids such as beclomethasone dipropionate (BDP) for the treatment of inflammatory lung diseases, e.g. asthma, chronic obstructive pulmonary disease and various type of pulmonary fibrosis. The thin layer evaporation method followed by sonication was used to prepare small non-ionic surfactant vesicles containing beclomethasone dipropionate. Light scattering experiments showed that beclomethasone dipropionate-loaded non-ionic surfactant vesicles were larger than unloaded ones and showed a significant (P<0.001) decrease of the zeta potential. The morphological analysis, by freeze-fracture transmission electron microscopy, showed the maintenance of a vesicular structure in the presence of the drug. The colloidal and storage stability were evaluated by Turbiscan Lab Expert, which evidenced the good stability of BDP-loaded non-ionic surfactant vesicles, thus showing no significant variations of mean size and no colloidal phase segregation. Primary human lung fibroblast (HLF) cells were used for in vitro investigation of vesicle tolerability, carrier-cell interaction, intracellular drug uptake and drug-loaded vesicle anti-inflammatory activity. The investigated NSVs did not show a significant cytotoxic activity at all incubation times for concentrations ranging from 0.01 to 1 μM. Confocal laser scanning microscopy showed vesicular carrier localization at the level of the cytoplasm compartment, where the glucocorticoid receptor (target site) is localized. BDP-loaded non-ionic surfactant vesicles elicited a significant improvement of the HLF intracellular uptake of the drug with respect to the free drug solution, drug/surfactant mixtures and empty vesicles used as references. The treatment of HLF cells with BDP-loaded non-ionic surfactant vesicles determined a noticeable increase of the drug anti-inflammatory activity by reducing the secretion of both constitutive and interleukin-1β-stimulated nerve growth factor (as inflammatory index) of 68% and 85%, respectively. Obtained data indicate that the investigated NSVs represent a promising tool as a pulmonary drug delivery system.

Scleroglucan: A Versatile Polysaccharide for Modified Drug Delivery

Scleroglucan is a natural polysaccharide, produced by fungi of the genus Sclerotium, that has been extensively studied for various commercial applications (secondary oil recovery, ceramic glazes, food, paints, etc.) and also shows several interesting pharmacological properties. This review focuses its attention on the use of scleroglucan, and some derivatives, in the field of pharmaceutics and in particular for the formulation of modified-release dosage forms. The reported investigations refer mainly to the following topics: natural scleroglucan suitable for the preparation of sustained release tablets and ocular formulations; oxidized and crosslinked scleroglucan used as a matrix for dosage forms sensitive to environmental conditions; co-crosslinked scleroglucan/gellan whose delivery rate can be affected by calcium ions. Furthermore, a novel hydrogel obtained with this polysaccharide and borate ions is described, and the particular structure of this hydrogel network has been interpreted in terms of conformational analysis and molecular dynamics. Profound attention is devoted to the mechanisms involved in drug release from the tested dosage forms that depend, according to the specific preparation, on swelling and/or diffusion. Experimental data are also discussed on the basis of a mathematical approach that allows a better understanding of the behavior of the tested polymeric materials.

Molecularly Imprinted Polymers for 5-Fluorouracil Release in Biological Fluids

The aim of this work was to investigate the possibility of employing Molecularly Imprinted Polymers (MIPs) as a controlled release device for 5-fluorouracil (5-FU) in biological fluids, especially gastrointestinal ones, compared to Non Imprinted Polymers (NIPs). MIPs were synthesized using methacrylic acid (MAA) as functional monomer and ethylene glycol dimethacrylate (EGDMA) as crosslinking agent. The capacity of the polymer to recognize and to bind the template selectively in both organic and aqueous media was evaluated. An in vitro release study was performed both in gastrointestinal and in plasma simulating fluids. The imprinted polymers bound much more 5-Fu than the corresponding non-imprinted ones and showed a controlled/sustained drug release, with MIPs release rate being indeed much more sustained than that obtained from NIPs. These polymers represent a potential valid system for drug delivery and this study indicates that the selective binding characteristic of molecularly imprinted polymers is promising for the preparation of novel controlled release drug dosage form.

Preparation and Characterization of Novel Gellan Gum Hydrogels Suitable for Modified Drug Release

Innovative hydrogels obtained by physical and chemical crosslinking of deacylated Gellan gum have been characterized in terms of water uptake, rheological properties and compressibility, and the behaviour of the tested materials, according to the type of the obtained network, is thoroughly discussed. The release from the various gels of loaded model molecules of different steric hindrance was also investigated and the trend of the release profiles has been related to the structures proposed for the physical and the chemical hydrogel.

Drug delivery strategies using polysaccharidic gels

Hydrogels are hydrophilic polymeric networks, with chemical or physical crosslinks, that are capable of swell and can retain a large amount of water. Among the numerous types of macromolecules that can be used for hydrogel formation, polysaccharides show very attractive advantages in comparison to synthetic polymers. They are widely present in living organisms, are usually abundant and show a number of peculiar physicochemical properties; furthermore, these macromolecules are, in most cases, non-toxic, biocompatible and can be obtained from renewable sources. For these reasons, polysaccharides seem to be particularly suitable for different applications in the wide field of pharmaceutics. As examples of the studies that have been carried out on this topic, this review will focus on two polysaccharides, alginate and xyloglucan. Alginate has been, and still is, extensively investigated and has numerous industrial applications, whereas xyloglucan was chosen because, although it has been much less studied, it shows interesting properties that should find important practical uses in the near future. The possible advantages of physical gels over those that are chemically crosslinked are also discussed.

In situ forming IPN hydrogels of calcium alginate and dextran-HEMA for biomedical applications

In situ forming hydrogels, which allow for the modulation of physico-chemical properties, and in which cell response can be tailored, are providing new opportunities for biomedical applications. Here, we describe interpenetrating polymer networks (IPNs) based on a physical network of calcium alginate (Alg-Ca), interpenetrated with a chemical one based on hydroxyethyl-methacrylate-derivatized dextran (dex-HEMA). IPNs with different concentration and degree of substitution of dex-HEMA were characterized and evaluated for protein release as well as for the behavior of embedded cells. The results demonstrated that the properties of the semi-IPNs, which are obtained by dissolution of dex-HEMA chains into the Alg-Ca hydrogels, would allow for injection of these hydrogels. Degradation times of the IPNs after photocross-linking could be tailored from 15 to 180 days by the concentration and the degree of substitution of dex-HEMA. Further, after an initial burst release, bovine serum albumin was gradually released from the IPNs over approximately 15 days. Encapsulation of expanded chondrocytes in the IPNs revealed that cells remained viable and, depending on the composition, were able to redifferentiate, as was demonstrated by the deposition of collagen type II. These results demonstrate that these IPNs are attractive materials for pharmaceutical and biomedical applications due to their tailorable mechanical and degradation characteristics, their release kinetics and biocompatibility.

A novel co-crosslinked polysaccharide: studies for a controlled delivery matrix

The formulation of a new controlled delivery system, based on a novel type of matrix obtained by the chemical reaction carried out in an aqueous medium on a mixed physical gel of gellan and scleroglucan, is described in this paper. The preparation yielded a new co-crosslinked polysaccharide (CCP) hydrogel, bearing carboxylic groups, that showed a sustained release behaviour that can be modulated by means of calcium ions. For the characterization of CCP, diffusion experiments through the swelled hydrogel were carried out in different environmental conditions and the release from tablets prepared with CCP and a model drug was evaluated. The addition of CaCl2 in the formulation of the dosage forms allowed a further marked reduction in delivery rate to be obtained; this effect is to be related to the free ionized carboxylic groups still present in the gellan moiety of CCP. The different behaviour of Ca+2 and Na+ ions is discussed.